NMR Screening for TRE Transcription Factor Aggregation
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Solution Overview
Problem
Current methods are inadequate for effectively screening active agents that modulate protein structure changes responsible for protein aggregate formation in TRE-based diseases, due to limited structural insights into homorepeat-containing proteins.
Innovation Solution
An in vitro method using Nuclear Magnetic Resonance (NMR) spectroscopy to detect incipient disordered protein structures triggered by TRE mutations, combined with light scattering procedures, to identify active agents that inhibit protein aggregation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If NMR spectroscopy is used to detect incipient disordered protein structures, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent segments the complex NMR spectroscopy platform into modular experimental components (HSQC, NOESY, ROESY experiments) that can be selectively applied based on specific detection needs, reducing overall system complexity while maintaining high measurement precision for detecting alternative protein structures
Solution Approach 2:
The patent introduces computational analysis tools as intermediaries between the NMR spectroscopy measurements and the detection of alternative protein structures, enabling precise identification of structural changes without requiring direct complex hardware modifications
2Measurement precision
If light scattering procedures are combined with NMR spectroscopy to detect protein aggregation, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent merges light scattering procedures with NMR spectroscopy into an integrated screening platform where each technique compensates for the other's limitations, achieving superior detection precision for protein aggregation events while sharing common sample handling infrastructure to mitigate complexity increases
Solution Approach 2:
The combined platform is designed with multi-functional capabilities where the same instrument system can perform both NMR spectroscopy and light scattering measurements, allowing a single device to serve multiple detection purposes and reducing overall system complexity
3Reliability
If screening is performed on full-length transcription factors, then reliability of detecting aggregation-prone structures is improved, but productivity decreases
Solution Approach 1:
The patent segments the full-length transcription factor into smaller functional domains or fragments for initial screening, allowing parallel processing of multiple segments that collectively cover the entire protein, thereby maintaining detection reliability while significantly increasing screening throughput
Solution Approach 2:
The patent applies partial action by focusing NMR spectroscopy detection on specific regions or residues most prone to aggregation rather than analyzing the entire protein uniformly, enabling faster screening while maintaining reliability in detecting aggregation-prone structures through targeted observation
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method allows for the precise detection of alternative protein structures responsible for aggregation, enabling the identification of active agents that block protein aggregation and potentially treat TRE-based diseases.
Implementation Method 1
determining by NMR spectroscopy any change in secondary structure in a transcription factor
Implementation Method 2
These low-populated, alternative structures are highly dynamic and can only be detected by solution NMR spectroscopy. This NMR methodology, in combination with light scattering procedures demonstrate that the alternative structures are responsible for protein aggregation
Data Source
Figure 1A~1B
Figure 1C
Figure 1D~1F
AI summary
The present invention refers to an in vitro method for the screening of active agents that inhibit changes in secondary structure in a transcription factor, wherein the changes in secondary structure are responsible for the formation of aggregates of said transcription factor, and wherein the method is based on NMR. The invention also refers the use of a transcription factor that comprises an amino acid repeat mutation, or a fragment thereof, for the in vitro screening of an active agent that inhibits changes in secondary structure in the transcription factor by NMR.